Project Summary How cancer cells co-evolve with their host microenvironment immune cells remains a critical question in immunotherapy. The Wu lab focuses on the study of antigen-specific interactions and their involvement in the cellular and molecular pathways underlying anti-tumor immunity. The DFCI Translational Immunogenomics Laboratory (TIGL) was created to expand technology and computational capabilities developed within the Wu Lab and provide a dedicated resource for immunogenomic analysis across different cancer types. My current portfolio of projects is focused on designing the development of single cell/genomics techniques to understand the crosstalk between tumor cells and immune cell populations within the tumor microenvironment and the selective pressure they mutually exert on each other. In particular: (i) I have developed the sensitive and robust rhTCRseq to determine paired alpha and beta-TCR clonotypes in single cells as well as to perform TCR repertoire analysis in bulk RNA samples. It has already revealed novel insights regarding the impact of immunotherapies, such as cancer vaccines, on the generation of antigen-specific immunity, and is being widely applied to the monitoring of other immunotherapy trials. (ii) For plate-based single-cell analysis and for bulk RNA samples, I have developed a novel strategy to perform qPCR and transcriptome sequencing. In addition, I have implemented a droplet-based 10x Genomics platform to study single cell transcriptome and surface protein expression in the same single cells. (iii) I have expertise in generating targeted multiplex RNA assays, aimed at simultaneous detection of genotype and transcriptome information at the single cell level. Altogether these diverse approaches have been and are being applied to several NCI-funded projects in the Wu Lab. Our new initiatives focus on the further development and implementation of novel single cell genomics technologies and their applications.